Holmila Reetta, Husgafvel-Pursiainen Kirsti
Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, FI-00250 Helsinki, Finland.
Cancer Detect Prev. 2006;30(1):1-6. doi: 10.1016/j.cdp.2005.10.004. Epub 2006 Feb 3.
Automated capillary electrophoresis single strand conformation polymorphism (CE-SSCP), denaturant gradient gel electrophoresis (DGGE), and direct sequencing were compared to investigate the benefits and sensitivity of each of the methods for detection of unknown TP53 mutations in human lung cancer.
Twenty previously analyzed DNA samples from lung tumors were examined under dummy laboratory codes for occurrence of mutations of the TP53 gene.
Mutations were found in 17 samples; 15 (88%) of them were detected by DGGE, 16 (94%) by CE-SSCP and 12 (71%) by direct sequencing. One of the two mutations that remained undetected in DGGE was in fact outside the sequence area covered by DGGE screening, thus rendering DGGE and CE-SSCP equally efficient in mutation detection. Direct sequencing performed less well in finding mutations than the two other assays, as also shown previously.
The study showed that CE-SSCP is a fast and highly reproducible method, which is considerably less laborious compared to DGGE, for screening of unknown TP53 mutations.
比较了自动毛细管电泳单链构象多态性（CE-SSCP）、变性梯度凝胶电泳（DGGE）和直接测序法，以研究每种方法检测人肺癌中未知TP53突变的优势和灵敏度。
对20份先前分析过的肺肿瘤DNA样本进行检测，样本采用虚拟实验室编码，以检测TP53基因的突变情况。
在17份样本中发现了突变；其中15份（88%）通过DGGE检测到，16份（94%）通过CE-SSCP检测到，12份（71%）通过直接测序检测到。DGGE未检测到的两个突变中有一个实际上在DGGE筛查覆盖的序列区域之外，因此DGGE和CE-SSCP在突变检测方面同样有效。如先前所示，直接测序在发现突变方面的表现不如其他两种检测方法。
该研究表明，CE-SSCP是一种快速且高度可重复的方法，与DGGE相比，在筛查未知TP53突变时工作量要小得多。
